Laser-Excited High-Sensitice Fluorescence Microscope System for Biological Cells using High-Speed Multi-Image Processing

本课题旨在研制一种高灵敏度的荧光显微镜系统，利用激光和多种图像处理方法等光电技术，对单个细胞内的荧光或散射强度分布进行定量测量，首先研制了一种以Ar激光为激发源的激光激发荧光显微镜系统，用于灵敏检测的SIT相机和用于图像信号的数字处理的图像处理器。实践证明，该系统可以检测含有某些有机染料的生物细胞的荧光分布，并对SIT相机的图像信号进行定量分析。基于上述研究的结果和理解，我们进一步改进了系统，采用图像增强器和新的图像处理器，以实现优异的高灵敏度以及更好的空间和时间分辨率。因此，来自硒的非常弱的荧光强度分布 关于我们 用该系统测量并三维显示含有临床上感兴趣的有机染料的几种细胞，其指示这些染料在细胞中的累积部分。荧光强度分布的时间变化，这主要是由光化学效应引起的也被检测到在短时间内不到34秒的定量间隔。此外，一个激光激发的高速，时间分辨荧光显微镜系统是新开发的研究在纳秒时间尺度的生物细胞中的各种染料的荧光衰减。利用该系统结合紫外脉冲<N_2>激光，首次对培养的癌细胞和溶液中的光敏性血卟啉和脱镁叶绿酸a的特性进行了测量和比较，并讨论了它们的不同行为，从而成功地构建了一种新型的利用高速多图像处理技术的单细胞激光激发高速荧光显微系统。该系统和研究成果在医学、生命科学和生物技术等领域具有重要的应用价值。少

This project aimed to the new development of high-sensitive fluorescence microscope system, which enables us to measure quntitatively fluorescence or scattering intensity distribution in a single viological cell utilizing optoelectronic technique including lasers and multiple image processing methods.At first, We developes a laser-excited fluorescence microscope systen using an Ar laser for excitation, a SIT camera for sensitive detection and an image processor for digital processing of image signals. It was proved that this system can detect the fluorescence distribution from biological cell containing some organic dyes and analyze quantitatively the image signal from the SIT camara. Based on the results and understandings derived in the above study, we further improved the system employing an image intensifier and a new image processor to realize excellent high-sensitivity as well as better spatial and temporal resolutions. Thus, very weak fluorescence intensity distributions from se … More veral kinds of cells containing clinically interesting organic dyes were measured and displayed three-dimensionally with this system, which indicate accumulated portions of these dyes with in the cells. Temporal changes in fluorescence intensity distribution, which were mainly caused by the photochemical effect were also detected quatitatively in a short time interval less than 34 sec.Moreover, a laser-excited high-speed, time-resolved fluorescrnce mivroscope system was newly developed to investigate fluorescence decays from various dyes in biological cells in nanosecond time scale. Using this system incorporating an UV pulsed <N_2> laser, we performed characteristics measurement and comparison of photosensitizing hematoporphyrin species and pheophorbide a in cultured cancer cells and solutions for the first time and their different behaviors were discussed.In summary, we have succeeded in constructing a novel type of laser-excited high-speed fluorescnce microscope system for single biological cells using highspeed multi-image processing technique. The system and the results achieved by this Research Project should be useful and valuable in the fields of medicine, lifescience and viotechnology and their practical applications are expected in the near future. Less